同种异体骨髓间充质干细胞穴位移植对梗死心肌及周围区细胞凋亡及相关蛋白的影响

背景：骨髓间充质干细胞移植途径主要包括外周静脉注射移植、冠状动脉移植以及心肌内注射移植，但存在迁移缺少靶向性，价格高昂等局限性。 目的：观察兔骨髓间充质干细胞穴位注射对梗死区心肌细胞凋亡及凋亡调控蛋白Fas,FasL,Bcl-2表达的影响。 设计、时间及地点：随机对照动物实验，于2007-06/08在辽宁中医药大学针灸系电生理实验室完成。 材料：抽取健康日本大耳白兔骨髓，分离培养骨髓间充质干细胞并传代；结扎日本大耳白兔左室支建立急性心肌梗死模型。 干预：将造模成功的日本大耳白兔30只随机分为3组，模型对照组不干预；穴位注射干细胞组于急性心肌梗死形成后1周，取心俞（双侧）、至阳、膻中4穴，穴位注射干细胞混悬液，1×l06细胞/穴；穴位注射生理盐水组在相同部位穴位注射等量生理盐水。以10只未造模兔为正常对照组。 主要观察指标：干预后第5周取心脏，TUNEL染色观察梗死区心肌细胞凋亡；免疫组化观察心肌Fas，FasL及Bcl-2蛋白的表达。 结果：模型对照组和穴位注射生理盐水组梗死区心肌细胞凋亡高于正常对照组和穴位注射干细胞组（P < 0.001，0.05）。穴位注射干细胞组Fas和FasL蛋白的表达明显低于模型对照组（P < 0.001），模型对照组则高于正常对照组（P < 0.05，0.001）。穴位注射干细胞组及正常对照组Bcl-2蛋白表达明显高于模型对照组和穴位注射生理盐水组（P < 0.001）。 结论：穴位移植骨髓间充质干细胞可使梗死及周围区心肌细胞凋亡数减少，其机制可能是通过Fas、FasL蛋白的减少、Bcl-2蛋白的增多来调节的。

Effects of allogenic bone marrow mesenchymal stem cell acupoint transplantation on cell apoptosis and related protein in infarcted myocardium and surrounding regions

BACKGROUND: Bone marrow mesenchymal stem cell (BMSCs) transplantation included peripheral vein injection transplantation, coronary artery transplantation and intramuscular injection transplantation. However, the migration lacks of target and the price is very high. OBJECTIVE: To investigate the effect of rabbit BMSCs acupoint injection on infarcted myocardium apoptosis and expressions of apoptosis-regulating proteins of Fas, FasL and Bcl-2. DESIGN, TIME AND SETTING: This randomized control animal experiment was performed at the Acupuncture electrophysiology laboratory of Liaoning University of Traditional Chinese Medicine from June to August 2007. MATERIALS: Bone marrow was collected from healthy Japanese rabbits. BMSCs were harvested and cultured. Acute myocardial infarction models were established by ligating left ventricles of Japanese rabbits. INTERVENTION: Thirty myocardial infarction rabbit models were randomly divided into 3 groups. Rabbits in the model control group were left intact. Stem cell suspension (1×l06 cells/per point) was injected into the Xinshu (BL 15) (bilateral), Zhiyang (DU 9) and Danzhong (RN 17) in the point injection stem cell group at 1 week after acute myocardial infarction. The same volume of saline was injected into the above mentioned points in the point injection saline group. Ten non-established model rabbits served as normal controls. MAIN OUTCOME MEASURES: Hearts were obtained at 5 weeks after intervention. Myocardium apoptosis was observed at infracted regions by TUNEL staining. Fas, FasL and Bcl-2 protein expression was detected in myocardium by immunohistochemistry. RESULTS: Myocardium apoptosis in the model control group and point injection saline group were higher than in the normal control group and point injection stem cell group (P < 0.001, 0.05). The expressions of Fas and FasL protein were significantly lower in the point injection stem cell group than in the model control group (P < 0.001), and higher in the model control group than in the normal control group (P < 0.05, 0.001). The expression of Bcl-2 protein in the point injection stem cell group and normal control group was significantly higher than in the model control group and point injection saline group (P < 0.001). CONCLUSION: Point transplantation of bone marrow mesenchymal stem cells can reduce the number of cardiomyocyte apoptosis in infarct zone, which may be adjusted through the reduction of Fas and FasL protein and the increasing of Bcl-2.